Additional file 1 of Engineered circular guide RNAs boost CRISPR/Cas12a- and CRISPR/Cas13d-based DNA and RNA editing

Additional file 1: Fig. S1 Establishment of the dLbCas12a-p300knock-in HEK293T cell line. Fig. S2 Circular gRNAs increase thetranscription efficiency of LbCas12a-based activators. Fig. S3 Establishment of the LbCas12a knock-in HEK293Tcell line and off-target analysis of LbCas12a with different gRNAs. Fig. S4 Circular gRNAs improve the DNA cleavage efficiency of LbCas12ain MCF7 cells. Fig. S5 comparison between cgRNA with other extendedstructuralized gRNAs. Fig. S6 Circular gRNAs improve the gene expressionor DNA cleavage of AsCas12a-based effectors. Fig. S7 Multiplexedgene activation and cleavage guided by cgRNAs. Fig. S8 Efficient andspecific RNA cleavage activity of CasRX with cgRNA. Fig. S9 Thetrans-cleavage activity of CasRx–mediated exogenous transcripts degradation. Fig.S10 The trans-cleavage activity of CasRx–mediated endogenous transcriptsdegradation. Fig. S11 The structures of circular gRNAs for LbCas12awith different linkers targeting IL1RN predicted by mFold. Fig. S12The structures of circular gRNAs for LbCas12a with different linkerstargeting IL1RN predicted by mFold. Fig. S13 The structures ofcircular gRNAs for CasRx with different linkers targeting STAT3predicted by mFold.

附加文件1：
图S1 dLbCas12a-p300敲入HEK293T细胞系的构建。
图S2 环状向导RNA（circular guide RNA，cgRNA）提升基于LbCas12a的转录激活因子的转录效率。
图S3 LbCas12a敲入HEK293T细胞系的构建及不同向导RNA介导的LbCas12a脱靶效应分析。
图S4 环状向导RNA可提升MCF7细胞中LbCas12a的DNA切割效率。
图S5 环状向导RNA与其他延伸型结构化向导RNA的对比分析。
图S6 环状向导RNA可提升基于AsCas12a的效应工具的基因表达或DNA切割能力。
图S7 环状向导RNA介导的多重基因激活与切割。
图S8 结合环状向导RNA的CasRX展现高效且特异性的RNA切割活性。
图S9 CasRx介导的外源转录本降解的反式切割活性。
图S10 CasRx介导的内源转录本降解的反式切割活性。
图S11 基于不同连接臂、靶向IL1RN的LbCas12a环状向导RNA的mFold预测结构。
图S12 基于不同连接臂、靶向IL1RN的LbCas12a环状向导RNA的mFold预测结构。
图S13 基于不同连接臂、靶向STAT3的CasRx环状向导RNA的mFold预测结构。